Bacillus popilliae cry18Aa operon is transcribed by sigmaE and sigmaK forms of RNA polymerase from a single initiation site.
Bacillus popilliae is an obligate pathogen for larvae of the insect family Scarabaeidae (Coleoptera). It forms parasporal crystals upon sporulation. The gene cry18Aa coding for the parasporal crystal protein and an upstream open reading frame, orf1, were previously isolated from B.popilliae. Here we report an analysis of cry18Aa transcription in Bacillus thuringiensis. The only transcriptional start site of cry18Aa was found 29 bp upstream of the open reading frame orf1, suggesting that orf1 and cry18Aa are transcribed as an operon. lacZ fusion to the cry18Aa promoter was used to follow the time-course of cry18Aa transcription in wild type B.thuringiensis and in various B.thuringiensis sporulation-deficient mutants (spo0A, sigE or sigK). In wild type B.thuringiensis, the cry18Aa promoter was activated 2 h after the end of exponential growth and the expression lasted to the late sporulation phase. The results of promoter activity in Spo+or Spo-backgrounds together with the results of primer extension experiments suggest that the transcription from this promoter can be driven by both sigmaE and sigmaK types of RNA polymerase at a single start site. The promoter region of cry18Aa operon fits the consensus sequences of both sigmaE and sigmaK dependent promoters of Bacillus.
Zhang
J
J
Zentrum für Molekularbiologie Heidelberg and Zoologisches Institut, Universität Heidelberg, Im Neuenheimer Feld 282, 69120 Heidelberg, Germany. zhang@sun0.urz.uni-heidelberg.de
Schairer
H U
HU
Schnetter
W
W
Lereclus
D
D
Agaisse
H
H
D000818:Animals; D019677:Artificial Gene Fusion; D001407:Bacillus / Q000235:genetics* / Q000472:pathogenicity; D001413:Bacillus thuringiensis / Q000235:genetics; D001427:Bacterial Toxins / Q000235:genetics; D001483:Base Sequence; D002874:Chromosome Mapping; D001517:Coleoptera / Q000382:microbiology; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D007763:Lac Operon; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D009876:Operon; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012808:Sigma Factor / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic
D001427:Bacterial Toxins; D017931:DNA Primers; D004269:DNA, Bacterial; D012808:Sigma Factor; D014157:Transcription Factors; C084190:sigma K; C063202:sporulation-specific sigma factors; D012321:DNA-Directed RNA Polymerases